Today I finished extracting the broodstock DNA You can see the details here!
I also helped Grace run her samples on the bioanalyzer at the end of the day.
Daily Archives: October 8
Yaamini’s Notebook: Gigas Broodstock DNA Extraction Part 9
Sample extraction: Day 2
Kaitlyn lead DNA extractions today while I bounced between lab, classes, and other obligations! She extracted DNA from the following samples:
Low pH:
- 1-T3
- 7-T2
- 8-T2
- 10-T3
- 3-T1
Ambient pH:
- UK-06
- 11-T4
- UK-05
- UK-08
- UK-02
Methods
Kailtyn and I followed this protocol. Here are some notes:
- Step 1: Kaitlyn carved out the tissue from histology blocks while I was in class! She scraped the tissue out for the first batch, so I know the technique between batches is consistent. She noted that some of the samples from this batch were shallower, so we may not be able to extract RNA from those samples.
Table 1. Mass of samples used for DNA extractions.
| Sample ID | Mass (mg) |
|---|---|
| 1-T3 | 19.7 |
| 7-T2 | 19.9 |
| 8-T2 | 19.8 |
| 10-T3 | 19.7 |
| 3-T1 | 20.2 |
| UK-06 | 20.0 |
| 11-T4 | 19.8 |
| UK-05 | 19.7 |
| UK-08 | 20.0 |
| UK-02 | 20.2 |
Figure 1. Histology blocks after carving out tissue.
- Step 9: Ethanol was evaporated out for 10 minutes
- Step 10: Kaitlyn noted we were low on Buffer TD1
- Step 15: For the first incubation-vortex set, the temperature was 56.9ºC instead of 56ºC. I loosened the foil and allowed some air flow, which brought the temperature to 56ºC.
- Step 17: Samples incubated an extra 3 minutes while waiting for the heat block to reach 80ºC
- Step 27: Samples were placed in the fridge for quantification on Tuesday
Results
Going forward
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